Inhibitory effect of puerarin on adipogenic differentiation of alcohol-induced human bone marrow mesenchymal stem cells

Yan YANG; Yali YU; Chunxi YANG; Xiaokun LI; Yisheng WANG; Yuebai LI

ABSTRACT

BACKGROUND:

Pueraria is a member of isoflavone characterizing by decreasing the toxic effects of alcohol,relieving the effect of alcohol withdrawal,anti-oxidative effect,removing oxygen free radicals,and preventing cell injury induced by lipid peroxidation.

OBJECTIVE:

To investigate the inhibitory effect of puerarin on adipogenic differentiation of alcohol-induced human bone marrow mesenchymal stem cells (hBMSCs).DESIGN,TIME AND

SETTING:

An in vitro observation based on cytology was performed at Laboratory of Biochemistry and Molecular Biology,Basic Medical College of Zhengzhou University in October 2008.MATERIALSThe bone marrow was taken from the adult healthy volunteers from the Department of Orthopedics of the First Affiliated Hospital of Zhengzhou University.Puerarin standard was provided by the National Institute for the Control of Pharmaceutical and Biological Products.

METHODS:

The primary hBMSCs were purified from the adult marrow by gradient centrifugation.The second passaged cells were divided into 3 groups.Blank control group Cells were cultured with DMEM culture media containing 10% fetal bovine serum;Model groupCells were cultured with 0.09 mol/L alcohol;Experimental groupCells were cultured with 0.09 mol/L alcohol and 10-6 mol/L puerarin;while,the culture liquid was changed every two or three days.Cells were cultured for 7 days in each group.MAIN OUTCOME

MEASURES:

PPARγ 2 gene expression with RT-PCR;amount of adipocyte with oil red "O" staining;alkaline phosphatase activity.

RESULTS:

PPAR-γ2 mRNA expression in the model group was significantly greater than experimental group and blank control group (P ＜0.05),while the expression in the experimental group was slightly greater than blank control group,but there was no significant difference (P ＞0.05).The number of adipocytes was the most in the model group,and the lipid droplet was large and abundant.The number of adipocytes in the experimental group was significantly less than model group (P ＜ 0.05),and there were few adipocytes in the blank control group.Compared with blank control group,alkaline phosphatase activity was decreased in the model group (P＜0.05);but the activity in the experimental group was increased compared to model group (P ＜ 0.05).

CONCLUSION:

Puerarin inhibits differentiation of hBMSCs into adipocytes induced by alcohol.